title: Targeting adenoviral vectors for enhanced gene therapy of uterine leiomyomas.
abstract: OBJECTIVE
  Is targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus -human somatostatin receptor subtype 2- arginine, glycine and aspartate-thymidine kinase), given in combination with ganciclovir (GCV) against immortalized human leiomyoma cells (HuLM) a potential therapy for uterine fibroids?
  
  
  CONCLUSIONS
  Ad-SSTR-RGD-TK/GCV, a targeted adenovirus, effectively reduces cell growth in HuLM cells and to a significantly greater extent than in human uterine smooth muscle cells (UtSM).
  
  
  BACKGROUND
  Uterine fibroids (leiomyomas), a major cause of morbidity and the most common indication for hysterectomy in premenopausal women, are well-defined tumors, making gene therapy a suitable and potentially effective non-surgical approach for treatment. Transduction of uterine fibroid cells with adenoviral vectors such as Ad-TK/GCV (herpes simplex virus thymidine kinase gene) decreases cell proliferation.
  
  
  METHODS
  An in vitro cell culture method was set up to compare and test the efficacy of a modified adenovirus vector with different multiplicities of infection in two human immortalized cell lines for 5 days.
  
  
  METHODS
  Immortalized human leiomyoma cells and human uterine smooth muscle cells were infected with different multiplicities of infection (MOI) (5-100 plaque-forming units (pfu)/cell) of a modified Ad-SSTR-RGD-TK vector and subsequently treated with GCV. For comparison, HuLM and UtSM cells were transfected with Ad-TK/GCV and Ad-LacZ/GCV. Cell proliferation was measured using the CyQuant assay in both cell types. Additionally, western blotting was used to assess the expression of proteins responsible for regulating proliferation and apoptosis in the cells.
  
  
  RESULTS
  Transduction of HuLM cells with Ad-SSTR-RGD-TK/GCV at 5, 10, 50 and 100 pfu/cell decreased cell proliferation by 28, 33, 45, and 84%, respectively (P < 0.05) compared with untransfected cells, whereas cell proliferation in UtSM cells transfected with the same four MOIs of Ad-SSTR-RGD-TK/GCV compared with that of untransfected cells was decreased only by 8, 23, 25, and 28%, respectively (P < 0.01). Western blot analysis showed that, in comparison with the untargeted vector Ad-TK, Ad-SSTR-RGD-TK/GCV more effectively reduced expression of proteins that regulate the cell cycle (Cyclin D1) and proliferation (PCNA, Proliferating Cell Nuclear Antigen), and it induced expression of the apoptotic protein BAX, in HuLM cells.
  
  
  CONCLUSIONS
  Results from this study need to be replicated in an appropriate animal model before testing this adenoviral vector in a human trial.
  
  
  CONCLUSIONS
  Effective targeting of gene therapy to leiomyoma cells enhances its potential as a non-invasive treatment of uterine fibroids.
authors.affiliation: Department of Obstetrics and Gynecology, Meharry Medical College, Nashville, TN 37208, USA.
authors.forename: S
authors.initials: S
authors.lastname: Nair
authors.forename: D T
authors.initials: DT
authors.lastname: Curiel
authors.forename: V
authors.initials: V
authors.lastname: Rajaratnam
authors.forename: C
authors.initials: C
authors.lastname: Thota
authors.forename: A
authors.initials: A
authors.lastname: Al-Hendy
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004274:DNA, Recombinant / Q000009:adverse effects / Q000378:metabolism* / Q000627:therapeutic use; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods; D005822:Genetic Vectors / Q000009:adverse effects / Q000378:metabolism* / Q000627:therapeutic use; D006801:Humans; D007889:Leiomyoma / Q000378:metabolism* / Q000628:therapy; D009215:Myometrium / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D014594:Uterine Neoplasms / Q000378:metabolism* / Q000628:therapy
chemical_list: D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D004274:DNA, Recombinant; D009363:Neoplasm Proteins
keywords: Ad-SSTR-RGD-TK; adenovirus vectors; gene therapy; uterine leiomyomas